Frozen Skin Tissue Block Preparation: A Protocol to Preserve Cutaneous Melanoma Samples from Murine Dorsal Skin

In This Article

Abstract

Source: Moon, H., et al. Spatial and Temporal Control of Murine Melanoma Initiation from Mutant Melanocyte Stem Cells. J. Vis. Exp. (2019)

This video describes the detailed protocol for isolating melanoma-containing murine dorsal skin, followed by cryo-embedding to prepare a frozen skin tissue block. The cryo-embedded tissue block can be sectioned and microscopically examined to study melanoma progression.

Protocol

All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Tissue Processing

Skin isolation
1. Obtain the following prior to starting: necropsy instruments, filter paper, and paper towels.
2. Euthanize the mice according to approved protocols, and confirm death prior to proceeding.
3. Using a hair clipper, shave any hair from the dorsal skin area. Lightly brush the area with a lint-free tissue to clear the area.
4. Make a small incision with sharp scissors just above the base of th....

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Results

Mouse skin sectioning process, cryomold use in diagram, tissue embedding in OCT medium.
Figure 1: Skin tissue isolation and cryo-embedding. Following euthanasia, shave the mouse dorsal skin region of interest and embed skin in cryomold containing OCT. The dotted line on the mouse represents the midline. Typically, 3 or 4 pieces of skin can be isolated .......